Objective To study the effects of human insulin-like growth factor 1(hIGF-1) gene transfection on the proliferation of NIH3T3 fibroblasts.Methods The plasmid of pcDNA3.1-hIGF-1 was transfected into NIH3T3 fibroblasts by using Lipofectin method.The positive cell clones were selected with G418 and cultured for 4 weeks.The stable expression of hIGF-1 in the positive cells was determined by in situ hybridization and immunocytochemical analysis.MTT assay and flow cytometer analysis were used to observe the proliferation of NIH3T3 fibroblasts.Results hIGF-1 mRNA and protein expressed in NIH3T3 fibroblasts transfected with pcDNA3.1-hIGF-1 by in situ hybridization and immunocytochemical analysis.MTT assay showed the A value of transfected NIH3T3 fibroblasts rose,compared with untransfected NIH3T3 fibroblasts group,the difference was significant(P

The incidence of systemic fungal infections have increased dramatically, moreover, drug resistance including either primary (intrinsic) or secondary (acquired) resistance, becomes one of the main reasons accounting for the failure of treating invasive fungal infections in the past decades. Nowadays, clinically available antifungal drugs are limited and their combination in antifungal therapy was not effective. It is expected to be a new strategy to synergistically sensitize antifungal drugs against drug-resistant fungi by using new small molecules. Based on the study in our research group and the reported work of others, we reviewed the research of the natural products which have synergistic effect with the antifungal agents against drug-resistant fungi. This review focused on the resource, structure, pharmacological activity, and action mechanism of the compounds, as well as somewhat in common, and would provide theoretical base for seeking new drug against drug-resistance fungi.
Antifungal Agents ; chemistry ; pharmacology ; Biological Products ; chemistry ; pharmacology ; Drug Synergism ; Fungi ; drug effects

Aim To investigate the effects of ursolic acid from loquat leaves on proliferation inhibition and expression of PPAR-γ 、TGF-β1 in rat hepatic stellate cells, so as to explore the mechanism of anti-hepatic fibrosis of UA.Methods HSC-T6 cells were randomly divided into blank group, rosiglitazone control group, the low, medium and high concentration of UA group to detect the cell proliferation inhibition by CCK-8 after 24,48,72 h.The content of type collagen Ⅰ in cell culture supernatant of each group was examined by ELISA.The expression of PPAR-γ mRNA, TGF-β1 mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR.Effects on HSC-T6 PPAR-γ and TGF-β1 protein from each group were detected by immunocytochemical method.Results CCK-8 results showed that the inhibitory rate of UA on cell proliferation increased with the prolongation of drug action time(P<0.01).ELISA results showed that with the increase of the concentration of UA, the content of type Ⅰ collagen content decreased(P<0.01).Real-time PCR results showed that with the increase of the concentration of UA, and the expression of PPAR-γ mRNA increased, the expression of TGF-β1 mRNA decreased(P<0.01).The results of immunocytochemistry showed that the expression of PPAR-γ protein was increased, and the expression of TGF-β1 protein was decreased with the increase of the concentration of UA(P<0.01).All effects mentioned above were dose-dependent.Moreover, the effects in the high concentration groups were stronger than those in control group.Conclusion UA can inhibit the proliferation of HSC-T6 cells, Which may be associated with the up-regulation of PPAR-γ expression and the down-regulation of TGF-β1 expression.

To establish a LC-MS/MS method for quantification of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in rats plasma and study its pharmacokinetics after administration of Mailuoning injection at a single dose to rats. Plasma samples were acidified with hydrochloric acid and extracted with ethyl acetate. The analytes were determined by LC-MS-MS using a ZOBAX SB C18 column with a mobile phase of methanol-water (containing 2 mmol x L(-1) ammonium acetic) (60:40)at a flow rate of 0.5 mL x min(-1) and detected using ESI with negative ionization mode. Ions monitored in the multiple reaction monitoring (MRM) mode were m/z 353.1/191.0 [M-H]- for chlorogenic acid, m/z 178.9/134.9 [M-H]- for caffeic acid, m/z 515.2/353.0 [M-H]-for 3,4-DCQA, m/z 193.0/133.9 [M-H]-for ferulic acid, m/z 146.9/102.9 [M-H]- for cinnamic acid and m/z 246.0/125.8 [M-H]- for tinidazole (IS). After administration of Mailuoning injection at a single dose to eight Sprague-Dawley rats, the concentrations of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in plasma were determined by LC-MS/MS method. The main pharmacokinetics parameters of measured data were caluculated by using DASver 1.0 software. The linear concentration ranges of the calibration curves for chlorogenic acid, caffeic acid, 3,4-DCQA and cinnamic acid were 2.006-1,027 microg x L(-1) (r = 0.999 6), 1.953-1,000 microg x L(-1) (r = 0.999 7), 28.51-1.459 x 10(4) microg x L(-1) (r = 0.998 9), 1.836-940.0, g x L(-1) (r = 0.997 7) and 4.780-2,447 microg x L(-1) (r = 0.998 6) respectively. The inner and inter-days relative standard deviations were both less than 5.0%, indicating legitimate precise and accuracy to the requirement of biological sample analysis. For chlorogenic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (49.78 +/- 12.81) min, (123.55 +/- 14.82) mg x min x L(-1) and (0.004 3 +/- 0.000 5) L x min(-1), respectively. For caffeic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (36.65 +/- 10.59) min, (91.67 +/- 11.77) mg x min L(-1) and (0.005 7 +/- 0.000 7) L x min(-1), respectively. For 3,4-DCQA, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (50.08 +/- 13.78) min, (278.34 +/- 31.82) mg x min x L-1 and (0.001 6 +/- 0.000 2) L x min(-1), respectively. For ferulic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (51.39 +/- 15.52) min, (34.72 +/- 4.67) mg x min x L(-1) and (0.000 4 +/- 0.0001) L x min(-1), respectively. For cinnamic acid, the pharmacokinetic parameter t1/2, AUCo-t, and CL were (74.42 +/- 18.32) min, (34.63 +/- 4.82) mg x min x L(-1) and (0.007 7 +/- 0.001 1) L x min-', respectively. The assay method is proved to be sensitive, accurate and convenient. It can be applied to the pharmacokinetic study of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid.
Animals ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Hydroxybenzoates ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

[ ABSTRACT] AIM:To observe the effect of high glucose on the protein expression of calreticulin ( CRT) and its association with cell apoptosis and mitochondrial dysfunction in the cardiomyocytes.METHODS: AC-16 cardiomyocytes were randomly divided into normal glucose group, high glucose group, high glucose+CRT siRNA group and isotonic con-trol group.The cell apoptotic rate, reactive oxygen species (ROS), mitochondrial membrane potential level, respiratory enzyme activity, and protein expression of CRT were observed.RESULTS: Compared with the cardiomyocytes in normal glucose group, the apoptotic rate and ROS production of cardiomyocytes increased in high glucose group, accompanying with the decreases in the mitochondrial membrane potential level and enzyme activitiy of the respiratory chain.The protein expression of CRT was significantly increased in high glucose group.However, compared with high glucose group, high glucose+CRT siRNA decreased the expression of CRT and attenuated the damage of mitochondria, but CRT siRNA did not reduce the ROS level in cardiomyocytes.CONCLUSION:High glucose brings about CRT over-expression to induce mito-chondrial injury, thus increasing myocardial apoptosis.

This study was aimed to select the best preparation form of Aurea Helianthus in order to provide a scientific basis for research and development of anti-myocardial ischemia drugs.The content of effective index components of preparation of Aurea Helianthus was detected using HPLC-PDA.Acute myocardial ischemia rat model introduced by injecting pituitrin was used to investigate the effect of anti-myocardial ischemia.The quality of preparation from Aurea Helianthus was characterized based on pharmacy and efficacy.And the characterized results were given correlation analysis.The results showed that the HPLC method for quantitative determination was verified by methodology.The average recoveries were in the range of 97.04％-102.44％.All three preparations of Aurea Helianthus had the efficacy of anti-myocardial ischemia in varying degrees.The enrichment had the highest effect with lowest extraction rate and the lowest dosage,which was the best form in medicinal-preparation application of Aurea Helianthus.It was concluded that the content of related drug components was higher with unique proportion in enrichment of Aurea Helianthus which was prepared based on drug system proved a high quality medicine.Therefore,the key to the improvement of efficacy was to explore the fittest proportion and improve the content of related pharmaceutical ingredients pertinently according to drug system.

Objective To implement visualization and surgical simulation of female pelvis based on virtual reality. Methods The successive cross-section images of pelvic part from a Chinese Visible Human (CVH) dataset were adopted. The important organs in pelvis were segmented initially. Then the VRML-based visualization and Dextrobeam-based virtual surgery simulation were carried out. Results A networked pelvic model and a surgical model were constructed with virtual reality techniques. The organs and structures in pelvis could be displayed clearly and the simulated surgical operations could be performed. Conclusion The application of virtual reality techniques can facilitate the anatomical teaching and surgery design. More significantly, a realistic practice platform will be provided for medical education and training.

Objective To investigate the characteristics of traditional Chinese medical syndrome types of fulminant and epidemic dengue fever patients admitted in Guangzhou and Xishuang banna in the year of 2013,and to ex plore the differences of etiology and pathogenesis, illness, and treatment for the patients in the two regions. Methods We collected the clinical data of 78 cases receiving integrative Chinese and western medicine from 255 patients admitted in Guangzhou Municipal Eighth People’s Hospital, and the clinical data of 39 cases receiving integrative Chinese and western medicine from 120 patients admitted in the People’s Hospital of Xishuangbanna Dai Nationality Autonomous Prefecture in the year of 2013. The traditional Chinese medical syndrome types and the syndrome scores of the total of 117 cases were investigated. The method of phenology was used for the analysis of epidemic time and epidemic region of dengue fever, and the theory of defense-qi-nutrient-blood syndrome differentiation of seasonal febrile diseases was used for the analysis of etiology and pathogenesis of dengue fever. Results ( 1) Dengue fever was epidemic in the first ten days of July and in the middle ten days of November of the year 2013 in Guangzhou region, and was epidemic in the middle ten days of August and the first ten days of October in Xishuangbanna region. The epidemicity of dengue fever in Guangzhou covered the end of summer and the whole autumn, and then disappeared before the coming of winter. In Xishuangbanna , the epidemicity of dengue fever was obvious in autumn, and disappeared in late autumn. ( 2) In the two hospitals, dengue fever patients were dominated by the syndromes of excessive heat in both Qifen and Xuefen, blood stasis blended with toxicity, excessive heat in Qifen, and lingering pathogens in order. (3) Before treatment, the scores of fever were higher in patients of Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital ( P<0.01) . After treatment for 3 days, fever scores were improved in both hospitals (P<0.05 or P<0.01), but the differences between the two hospitals were insignificant (P>0.05) . After treatment for 6 days, fever disappeared in patients of both hospitals. (4) Before treatment, the scores of syndromes were higher in patients of Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital ( P<0.05) . After treatment for 3 days, syndorme scores were improved in both hospitals ( P<0.01) , but the syndrome scores were higher in Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital. After treatment for 6 days, syndrome scores were much improved in patients of both hospitals compared with those after treatment for 3 days (P<0.01) . Conclusion In dengue fever patients admitted in Guangzhou and Xishuangbanna region, the syndrome of excessive heat in both Qifen and Xuefen is the leading type, and then comes blood stasis blended with toxicity. The illness state of patients in Guangzhou region is milder than that of the patients in Xishuangbanna region, the time for symptom relief is about one week, and similar therapeutic effect can be achieved in the two regions .

ObjectiveTo explore the effects of bone morrow stromal cells (BMSCs) on the neurological behavior of rats with traumatic brain injury (TBI).MethodsTwenty-four SD rats were randomly and equally divided into control group, TBI group and BMSC group. The weight-drop device was adapted to establish the TBI model. The injury severity and its outcome were evaluated by a set of criteria termed neurological severity score (NSS). Brain tissues were harvested at day 14 to observe the survival and migration of the transplanted cells.Bax expression was detected by RT-PCR. Results NSS was (12 ±3 ) points in the TBI group, significantly higher than (7 ± 1 ) points in the BMSC group (P ＜0.05). The transplanted BMSCs could survive and migrate. Moreover, BAX, a crucail apopotosis gene, was down-regulated to 0.9 ±0.1 in the BMSC group, compared with 1.1 ±0.2 in the TBI group (P ＜0.05). ConclusionsBMSC transplantation is available to improve the neurological function, as may be associated with the Bax.

ObjectiveTo investigate the role of reducing the door-to-needle time for patients with acute ischemic stroke based on the quality improvement program of PDCA cycle.MethodsConsecutive patients with acute ischemic stroke admitted to hospital were registered prospectively from January 1, 2016 to September 30, 2016.Questionnaires and time tracking method were used to investigate the door-to-needle (DNT) and its influencing factors.PDCA cycle method was used to improve the stroke channel workflow and the changing trend of DNT was analyzed.ResultsA total of 71 patients with acute ischemic stroke were enrolled.After 3 PDCA cycles, DNT (median, interquartile range) from 100.0 min (65.5-127.0 min) reduced to58.0 min (45.5-80.0 min) (Z=11.689, P<0.001), the proportion of the patients with DNT ≤60 min increased from 19.05% to 60.00% (χ2=7.893, P=0.019).Conclusions The quality improvement program of PDCA cycle may effectively reduce the time of DNT in patients with acute ischemic stroke.